Quality control method for commercially available wild Jujube leaf tea based on HPLC characteristic fingerprint analysis of flavonoid compounds

Flavonoids are the main active components of natural medicinal plants with many physiological functions. In this study, an HPLC fingerprinting method based on the distribution and relative amount of 11 bioactive flavonoids was established for the quality evaluation of commercially available wild Jujube leaf tea (JLT) from China. Separation of the crude flavonoid extract was achieved on a column filled with C₁₈ material with a high carbon content. The flavonoids in wild JLT were identified based on UV spectroscopy and accurate mass measurements by TOF‐MS. Twenty‐one batches of practical samples collected from different habitats were analyzed by using the developed HPLC method to construct the HPLC characteristic fingerprint of wild JLT. Then, combined with clustering and similarity analyses, the HPLC characteristic fingerprint was used for the authentication and quality evaluation of commercial wild JLT. Results indicated that the proposed HPLC characteristic fingerprint reflected the inherent characteristics of wild JLT collected from different regions. Authenticity identification and quality control of commercially available wild Jujube tea were achieved based on the HPLC characteristic fingerprint analysis. This new approach to bioactive component profiling provided a promising reference method for the quality evaluation of commercial wild flower and plant tea.     

Carbon molecular sieve based micro‐matrix‐solid‐phase dispersion for the extraction of polyphenols in pomegranate peel by UHPLC‐Q‐TOF/MS

A rapid, simple, and efficient sample extraction method based on micro‐matrix‐solid‐phase dispersion (micro‐MSPD) was applied to the extraction of polyphenols from pomegranate peel. Five target analytes were determined by ultra‐HPLC coupled with Q‐TOF/MS. Carbon molecular sieve (CMS) was firstly used as dispersant to improve extraction efficiency in micro‐MSPD. The major micro‐MSPD parameters, such as type of dispersant, amount of dispersant, grinding time, and the type and the volume of elution solvents, were studied and optimized. Under optimized conditions, 26 mg of pomegranate peel was dispersed with 32.5 mg of CMS, the grinding time was selected as 90 s, the dispersed sample was eluted with 100 μL of methanol. Results showed that the proposed method was of good linearity for concentrations of analytes against their peak areas (coefficient of determination r² > 0.990), the LOD was as low as 3.2 ng/mL, and the spiking recoveries were between 88.1 and 106%. Satisfactory results were obtained for the extraction of gallic acid, punicalagin A, punicalagin B, catechin, and ellagic acid from pomegranate peel sample, which demonstrated nice reliability and high sensitivity of this approach.     

Liquid chromatography coupled with time‐of‐flight tandem mass spectrometry for comprehensive phenolic characterization of pomegranate fruit and flower extracts used as ingredients in botanical dietary supplements

Pomegranate (Punica granatum L.) fruit and flower extracts, which are rich sources of bioactive phenolics, are widely utilized as ingredients in botanical dietary supplements. While the phenolic characterization of extracts of pomegranate fruit has been previously studied by liquid chromatography with tandem mass spectrometry, there is lack of similar data for pomegranate flowers. Herein, liquid chromatography with time‐of‐flight tandem mass spectrometry was utilized to comprehensively characterize the phenolics present in two pomegranate extracts, previously studied for their in vitro and in vivo biological effects, namely, a patented commercial pomegranate fruit extract (Pomella®), and a pomegranate flower extract. Seventy‐one phenolics were characterized in the pomegranate fruit extract with the vast majority identified in the flower extract. However, only the pomegranate fruit extract contained tannin‐glucuronides and two punicalagin isomers (a characteristic pomegranate phenolic) were identified in this extract while four were identified in the flower extract. The previously reported compounds, pomellatannin and punicatannins A/B, were identified as unique chemical markers in the pomegranate fruit and flower extracts, respectively. This study will aid in the quality control, authentication, and standardization of these botanical ingredients to evaluate their potential health benefits in future planned pre‐clinical and clinical studies. Also, this is the first phenolic characterization of a pomegranate flower extract using liquid chromatography with tandem mass spectrometry.     

Multiple compounds determination and fingerprint analysis of Lidanpaishi tablet and keli by high-performance liquid chromatography

The objective of this paper was to establish an efficient method for quality control of Lidanpaishi (LDPS) tablet and keli, two famous traditional Chinese medicines. A simple and reliable high-performance liquid chromatography (HPLC) coupled with photodiode array detector (DAD) method was developed both for fingerprint analysis (FA) and quantitative determination. In quantitative analysis, linear regressions, limit of detection (LOD) and quantification (LOQ), intra-day and inter-day precisions, recovery, repeatability and stability were all tested and good results were obtained to simultaneously determine the 15 marker compounds, namely chlorogenic acid, rhaponticin, 6,7-dimethoxycoumarin, naringin, hesperidin, neohesperidin, baicalin, wogonoside, baicalein, wogonin, chrysin, honokiol, magnolol, emodin, and chrysophanol in the herbal drugs. In fingerprint analysis, 34 peaks were selected as the characteristic peaks to evaluate the similarities of different samples collected from different pharmaceutical companies in China according to the State Food and Drug Administration (SFDA) requirements, and two kinds of data, relative retention time (RRT) and relative peak area (RPA) were used to identify the common peaks in 15 samples for investigation. Furthermore, hierarchical cluster analysis (HCA) was also performed to evaluate the variation of the herbal drugs. The present approach, i.e. HPLC coupled with multiple compounds determination (MCD) and FA is a powerful and meaningful tool to comprehensively conduct the quality control of traditional Chinese medicines.     

高效液相色谱测定石榴皮水提取物中4种多酚化合物的含量

建立了石榴皮水提取物中没食子酸、安石榴甙A、安石榴甙B和鞣花酸4种多酚化合物的高效液相色谱(HPLC)分析方法.样品经前处理并过滤后,采用HPLC检测,外标法定量.在最佳分析条件下,上述4种多酚化合物的线性范围分别为7.48 ～ 149.50、15.36～153.55、27.65 ～276.45、7.13 ～ 114....     

Pomegranate By-Products as Natural Preservative to Prolong the Shelf Life of Breaded Cod Stick

This work evaluated the efficacy of pomegranate byproducts, specifically peel powder, as valid preservatives for food quality. Ready-to-cook cod sticks breaded with pomegranate peel powder were prepared. Shelf-life tests were conducted on breaded cod sticks during refrigerated storage (17 days) at 4 °C, monitoring the pH, microbiological and sensory quality. In addition, the nutritional quality of both the breaded and control samples was assessed. The results highlighted that active samples showed higher phenol and flavonoid content and higher antioxidant activity compared to the control fish, suggesting that pomegranate peel powder was responsible for a significant increase in cod stick nutritional quality. Furthermore, the cod stick active breading led to a delay in microbial growth without affecting the sensory properties; rather, it helped slow down the sensory attribute decline during the refrigerated storage. The data suggest that using pomegranate byproducts in breaded cod stick was effective in prolonging its shelf life, as well as improving its nutritional quality. Therefore, pomegranate peel powder can be considered as a potential resource as natural food preservative.     

Fingerprint and multicomponent quantitative analysis for the quality evaluation of Sibiraea angustata leaves by HPLC-DAD and HPLC-ESI-MS/MS combined with chemometrics

Sibiraea angustata leaves, known as a traditional Tibetan medicine, have been specially used in the treatment of indigestion and obesity. In the study, a simple and sensitive high-performance liquid chromatography (HPLC) method with a diode array detector (DAD) was established to solve the problem of lacking quality standard of S. angustata leaves, including the fingerprint analysis and quantification of six characteristic components. The analytical method was validated for linearity, repeatability, stability, recovery, and specificity. Seventeen raw samples and 1 processed sample of S. angustata leaves were collected from different locations of China to establish the fingerprint. The chemometric methods, including similarity analysis (SA), principal component analysis (PCA), and hierarchical clustering analysis (HCA), were applied to distinguish the 18 batches of S. angustata samples. The results successfully sorted these samples into five clusters and kept in line with each other. According to the result of the fingerprint analysis, 21 peaks were extracted to be the common peaks and most of them were identified by mass spectrometry (MS) with electron-spray ionization (ESI) in the negative mode. Meanwhile, the loading plot of PCA further indicated that the peaks of neochlorogenic acid, chlorogenic acid, ferulic acid, rutin, hyperin, and isoquercitrin played a greater role in the discrimination among the 21 peaks. So the six components mentioned above were investigated as index constituents to evaluate the quality of S. angustata leaves from different locations. The study demonstrated that the developed new method was a beneficial approach for authentication and quality evaluation of S. angustata leaves.     

Chemical fingerprint analysis of Fritillaria delavayi Franch. by high-performance liquid chromatography

Bulbus of Fritillaria delavayi Franch. is the most commonly used antitussive and apophlegmatic in China and commonly prepared by water decoction. In this study, a novel and reliable method of high-performance liquid chromatography (HPLC) was developed both for quantitative analysis of 10 bioactive compounds (uracil, cytidine, inosine, uridine, guanosine, thymidine, adenosine, hypoxanthine, adenine and 2-deoxyadenosine) and chemical fingerprint analysis of F. delavayi Franch. In quantitative analysis, 10 compounds showed good regressions (R(2) > 0.9982) within test ranges and the recovery of the method was in the range of 96.33-104.51%. In the fingerprint analysis, 11 characteristic peaks were selected to evaluate the similarities of F. delavayi Franch. samples, and the HPLC chromatograms of 16 samples from different regions of China showed similar patterns. The results from the experiment demonstrated that the combinations of the quantitative and chromatographic fingerprint analysis offer an efficient way to evaluate the quality consistency of F. delavayi Franch.     

Influence of Cotton Pre-Treatment on Dyeing with Onion and Pomegranate Peel Extracts

In this paper the possibility of applying natural dyes on cellulose fibres were researched with respect to the impact of cotton material pre-treatment (scouring, chemical bleaching, mercerization and mordanting), using renewable sources of natural dyes (waste as a source). As mordants, metal salts of copper, aluminium and ferrum were used, and the influence on colour change as well as on fastness properties were analysed. The natural dyes were extracted from onion peel (Allium cepa L.) and pomegranate peel (Punica granatum L.). In spectrophotometric analysis performed of the plant extracts, the onion extract has peaks at 400 and 500 nm, resulting in red-orange colourations and the pomegranate extract shows a maximum at 400 nm, i.e., in the yellow region, which is characteristic of punicalin. Results show significant influence of cotton pre-treatments on colour appearance and fastness properties, caused by pre-treatments affecting the properties and structure of the cotton itself. The positive effect of mercerization on dye absorption and bonding is confirmed. For wash and light fastness properties, more satisfactory results have been obtained for yarns dyed with pomegranate peel natural dye, and the key importance of mordants for fastness properties has been confirmed.     

Comparison of the chromatographic fingerprint,multicomponent quantitation and antioxidant activity of Salvia miltiorrhiza Bge. between sweating and nonsweating

Salvia miltiorrhiza Bge. is a traditional Chinese medicine applied in the treatment of various diseases in clinical practice. In the course of its processing, S. miltiorrhiza Bge. is usually processed by sweating. This study employed 10‐component contents determination coupled with high‐performance liquid chromatography (HPLC) fingerprint and antioxidant activity to investigate the effect of sweating on S. miltiorrhiza Bge. so as to evaluate the quality of S. miltiorrhiza Bge. The HPLC method was performed using C₁₈ and 0.05% phosphoric acid aqueous solution–acetonitrile with a gradient elution system. It was validated for linearity, precision, repeatability, stability and recovery. Similarity analysis, principal components analysis and antioxidant activity assays were used to compare sweated S. miltiorrhiza Bge. (SSM) and nonsweated S. miltiorrhiza Bge. (NSSM). SSM and NSSM showed good similarities in HPLC fingerprint (>0.9), but principal components analysis could classify the HPLC fingerprint and 10‐component quantitation analysis. Meanwhile, the antioxidant activity of SSM was significantly higher than that of NSSM (p < 0.01). The results of this study indicated that sweating could alter the content of chemical constituents in S. miltiorrhiza Bge., and could also improve its antioxidant activity. In addition, the method not only affords a viable strategy for comparing SSM and NSSM and assessing the quality of S. miltiorrhiza Bge., but also provides a reference for other herbal medicine that suffers from sweating.     

仿刺参中皂苷类成分的高效液相色谱指纹图谱

利用高效液相色谱法建立了仿刺参皂苷类成分的指纹图谱,为仿刺参的质量控制提供了新的方法。采用固相萃取制备供试品溶液,选用Zorbox SB-C18色谱柱(250 mm×4.6 mm, 5 μm),以乙腈-0.1%磷酸水溶液为流动相进行梯度洗脱,检测波长为205 nm,柱温30 ℃。分析了不同产地的10批仿刺参样品,采用国家药典委员会推荐的“中药色谱指纹图谱相似度评价系统(2004 A版)”处理谱图,确定了6个共有峰。计算了10个样本间的指纹图谱相似度,所得相似度计算结果均大于0.97。该方法具有良好的稳定性和重现性,可用于仿刺参的质量控制。     

Chemical Fingerprint Analysis and Quantitative Analysis of Saccharides in Morindae Officinalis Radix by HPLC-ELSD

A method based on high performance liquid chromatography with evaporative light scattering detection (HPLC-ELSD) was developed for the quantitative analysis of three active compounds and chemical fingerprint analyses of saccharides in Morindae officinalis radix. Ten batches of Morindae officinalis radix were collected from different plantations in the Guangdong region of China and used to establish the fingerprint. The samples were separated with a COSMOIL Sugar-D column (4.6 mm × 250 mm, 5 μm) by using gradient elution with water (A) and acetonitrile (B). In addition, Trapped-Ion-Mobility (tims) Time-Of-Flight (tims TOF) was used to identify saccharides of Morindae officinalis radix. Fingerprint chromatogram presented 26 common characteristic peaks in the roots of Morinda officinalis How, and the similarities were more than 0.926. In quantitative analysis, the three compounds showed good regression (r = 0.9995–0.9998) within the test ranges, and the recoveries of the method were in the range of 96.7–101.7%. The contents of sucrose, kestose and nystose in all samples were determined as 1.21–7.92%, 1.02–3.37%, and 2.38–6.55%, respectively. The developed HPLC fingerprint method is reliable and was validated for the quality control and identification of Morindae officinalis radix and can be successfully used to assess the quality of Morindae officinalis radix.     

基于化学模式识别技术的枳实HPLC定量指纹图谱研究

建立了枳实的高效液相色谱(HPLC)指纹图谱分析方法.色谱柱为Tnature-ACCHROM C18色谱柱(4.6 mmx250 mm,5 μm);以乙腈-0.5％甲酸水溶液为流动相进行梯度洗脱,结合液相色谱-四极杆飞行时间质谱(HPLC-QTOF-MS)联用技术对枳实指纹图谱中的共有峰进行鉴定;采用相似度评价、聚类分...     

Chemical Fingerprint and Determination of Triterpenoids in Cultured Cyclocarya paliurus Cells by High-Performance Liquid Chromatography

Cyclocarya paliurus is a medicinal plant containing various bioactive components with significant health benefits. Cell cultures of C. paliurus have been used to produce these bioactive metabolites. A chemical fingerprint was obtained by high-performance liquid chromatography (HPLC) to monitor the synthesis of major triterpenoids in cultured C. paliurus cells and provide a reliable quality assessment for the cell strain screening. The determination of five triterpenoids, namely, maslinic acid, corosolic acid, betulinic acid, oleanic acid, and ursolic acid, was also performed. The HPLC method for the determination of triterpenoids in the cultured cells was accurate, stable, and reliable, and therefore suitable for chemical fingerprint analysis. Sixteen C. paliurus cell strains varied dramatically in their triterpenoid accumulations. The concentrations of the triterpenoids were 0.45–2.19 (maslinic acid), 0.92–5.34 (corosolic acid), 2.58–4.70 (betulinic acid), 4.07–12.47 (oleanic acid), and 12.64–40.98 (ursolic acid) mg/g. A high yield cell strain had a total triterpenoid concentration of 66.34?mg/g. Ten peaks in the HPLC chromatogram with reasonable height and high resolution were assigned as characteristic for fingerprint and analysis. A reference fingerprint was also obtained for cell strain assessment.     

Chromatographic fingerprint analysis and simultaneous determination of eight lignans in Justicia procumbens and its compound preparation by HPLC-DAD

HPLC fingerprints were developed for the quality evaluation of Justicia procumbens and its compound preparation, Jian-er syrup, together with the simultaneous quantification of eight arylnaphthalide lignans (6'-hydroxy justicidin B, 6'-hydroxy justicidin A, 6'-hydroxy justicidin C, justicidin B, chinensinaphthol methyl ether, justicidin C, taiwanin C, and neojusticin A). Samples were separated with a Shiseido Capcell Pak C(18) reversed-phase column (250×4.6 mm id, 5 μm) using acetonitrile and water as the mobile phase. The column temperature was maintained at 35°C and the wavelength of detector was set at 256 nm. For fingerprint analysis, 17 peaks were selected as the characteristic peaks for the evaluation of the similarities among different J. procumbens samples collected in different places. The structures of lignans were confirmed by diagnostic fragments in the positive ESI-MS(n) . The new method was successfully applied for the chromatographic fingerprint analysis and simultaneous determination of eight lignans in its compound preparation, Jian-er syrup. All the results indicated that HPLC fingerprint assay in combination with multi-marker determination afforded a useful method for the quality control of J. procumbens and its compound preparation, Jian-er syrup.     

Quality evaluation of Hpericum japomicum by using high‐performance liquid chromatography coupled with photodiode array detector and electrospray ionization tandem mass spectrometry

A high‐performance liquid chromatography coupled with photodiode array detection and electrospray ionization tandem mass spectrometry (HPLC‐PAD‐ESI‐MSⁿ) method was developed to evaluate the quality of Hpericum japomicum through establishing chromatographic fingerprint and simultaneous determination of seven phenolic compounds. The analysis was achieved on an Ultimate XB‐C₁₈ analytical column (250 mm × 4.6 mm i.d., 5 µm) using an aqueous solution of acetic acid (pH 3.8) and methanol as the mobile phase. Ten samples of H. japomicum from various habitats were investigated and the correlation coefficients of similarity were determined from the HPLC fingerprints. By using an online ESI‐MSⁿ, 20 common peaks in chromatographic fingerprints were identified as phenols, including flavones and their glycosides, flavonones and their glucosides, flavanols, xanthones, phloroglucinols, phenyl propanoids and chromones. Based on the above study, seven phenols which are considered to be major constituents in H. japomicum, including 3,4‐dihydroxybenzoic acid (1), taxfolin‐7‐O‐α‐l ‐rhamnoside (7), 7‐dihydroxy‐2‐(1‐methylpropyl)chromone‐8‐β‐d ‐glucoside (8), isoquercitrin (14), quercitrin (16), quercetin‐7‐O‐α‐l‐ rhamnoside (18) and quercetin (19) were quantified by the validated HPLC‐PAD method. This developed method by combination of chromatographic fingerprint and quantification analysis could be applied to control the quality of H. japomicum. Copyright © 2009 John Wiley & Sons, Ltd.     

Quality control of Citri reticulatae pericarpium: exploratory analysis and discrimination

Extracts of Citri reticulatae pericarpium (PCR) are commonly used in the Traditional Chinese Medicine. The quality control of PCR is currently performed by single marker analysis, which can hardly describe the complexity of such natural samples. In this study, a fingerprint methodology for PCR based on high-performance liquid chromatography (HPLC) was developed and validated. A total of 69 fingerprints of authenticated PCR samples, commercial PCR samples, mixed peel samples, and other Citrus peels were recorded. Exploratory data analysis allowed optimizing the extraction procedure and detecting mixed peel samples. Once the optimizations were performed and the method validated, discrimination between the authentic PCR samples and all other samples was performed by p-Discriminant Partial Least Squares. The established model was able to differentiate between classes with a high reliability for each sample. Furthermore, evaluation of the score and loading plots of the model indicated nobiletin, tangeretin, naringin and hesperidin as important markers for the quality control of PCR.     

Development of a novel method combining HPLC fingerprint and multi-ingredients quantitative analysis for quality evaluation of traditional Chinese medicine preparation

A novel method combining high performance liquid chromatography (HPLC) fingerprint and simultanous quantitative analysis of multiple acitve components was developed and validated for quality evaluation of one type of traditional Chinese medicine preparations: Shuang-huang-lian (SHL) oral liquid formulation. For fingerprint analysis, 45 peaks were selected as the common peaks to evaluate the similarities among several different SHL oral liquid preparations collected from manufacturers. Additionally, simultanous quantification of eleven markers, including chlorogenic acid, caffeic acid, rutin, forsythiaside, scutellarin, baicalin, forsythin, luteoloside, apigenin, baicalein and wogonin, was performed. Statistical analysis of the obtained data demonstrated that our method has achieved desired linearity, precision and accuracy. Finally, concentrations of these eleven markers in SHL oral liquid prepared by different manufacturers in China were determined. These results demonstrated that the combination of HPLC chromatographic fingerprint and simultaneous quantification of multi-ingredients offers an efficient and reliable approach for quality evaluation of SHL oral liquid preparations.     

Quality consistency evaluation of commercial transfer factor injections by chromatographic fingerprint combined with multivariate statistical analysis

The current quality control methods relying mainly on chromogenic reaction can hardly ensure the quality and safety of the biochemical drug with complex chemical composition. Therefore, a chromatographic fingerprint method was developed for the quality evaluation of a multicomponent biochemical drug, transfer factor injection. High‐performance liquid chromatography fingerprint was measured by using a C₁₈ column (250 × 4.6 mm, 5 µm) with a mobile phase composed of 0.1% trifluoroacetic acid–water and 0.085% trifluoroacetic acid–acetonitrile under gradient elution. The developed method was validated and was subsequently applied to 57 batches of commercial products which were sampled by National Drug Assessment Program. High‐resolution mass spectrometry analysis was performed on characteristic peaks of fingerprints, and a series of amino acids, nucleosides, and deoxynucleosides were identified. In the fingerprint assessments, principal component analysis and Hotelling T² analysis yielded the best results. The results generally indicated that there was a significant difference among products of batch‐to‐batch or from different manufacturers. Abnormal samples and its discriminatory components were also explored. In summary, the established fingerprinting method with multivariate statistical analysis could offer an efficient, reliable, and practical approach for quality consistency evaluation of transfer factor injection, providing a reference for the quality control of other multicomponent biochemical drugs.     

The difference between blood‐associated and water‐associated herbs of Danggui‐Shaoyao San in theory of TCM,based on serum pharmacochemistry

Danggui‐Shaoyao San (DSS) is a famous Chinese formula for activating blood circulation and promoting urination. This study was to investigate the difference of material basis between a blood‐associated herbs group and a water‐associated herbs group. According to the theory of traditional Chinese medicine, the formula can be divided into a blood‐associated herbs group (Angelica sinensis, Paeonia lactiflora and Ligusticum chuanxiong) and a water‐associated herbs group (Atractylodes macrocephala, Alisma orientale and Poria cocos). The HPLC fingerprint of the formula was established for quality control. Serum samples from rats, orally administrated DSS, and the decomposed recipes of DSS, were analyzed by HPLC‐DAD and the transitional blood components of DSS were identified. Twenty‐one common peaks were identified in the fingerprint of DSS. Contents of paeoniflorin, albiflorin, ferulic acid and alisol B 23‐acetate in co‐decoction were significantly higher than those in individual decoction. Eleven peaks belonged to the blood‐associated herbs group (four metabolites and seven prototype components; paeoniflorin and ferulic acid appeared in prototype components), whereas six peaks belonged to the water‐associated herbs group (three metabolites and three prototype components). It was concluded that the serum pharmacochemistry is a meaningful approach for clarifying the difference between blood‐associated and water‐associated herbs in chemical composition. Copyright © 2015 John Wiley & Sons, Ltd.